高通量测序分析臭豆腐中的细菌菌种多样性
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摘要
通过高通量测序的方法以及传统分离培养方法对臭豆腐中的细菌的微生物群进行分析。通过传统培养的方法分离到49株细菌,经鉴定隶属于9个属15个种。其中有乳酸菌11种,其它细菌4种。分离鉴定到属水平的细菌中,属于明串珠菌属的菌株最多。高通量测序结果显示,在样品中乳酸菌占有较高的比例,H1和H3样品中超过50%的细菌属于乳酸菌。高通量测序共检测到212种细菌,其中66种属于乳酸菌。臭豆腐的菌群构成十分复杂,暂时没有一种菌种是臭豆腐菌群中的优势菌种。沙克乳杆菌在本次研究分离的样品中所占比例相对较高,在每个样品中都能分离到。
The aim of this study was to investigate the microbiota of bacteria in stinky tofu samples through highthroughput sequencing and culture dependent. Forty-nine strains were isolated and identified as 9 genera and 15 species,with 11 of which were lactic acid bacteria and 4 as other bacteria. Particularly, Leuconostoc bacteria turned out to be the majority of the isolated microbe. On the other hand, data from high-throughput sequencing revealed that more than 50% of the bacteria belong to lactic acid bacteria(LAB) in H1 and H3 samples. A total of two hundred and twelve species were found by high-throughput sequencing and sixty-six of them belonged to LAB. High-throughput sequencing indicated that stinky tofu flora composition is very complicated and no one species was identified as the dominant strain among all the stinky tofu in this study. Lactobacillus sakei is isolated from each sample and takes a relatively high percentage in this specific niche.
The aim of this study was to investigate the microbiota of bacteria in stinky tofu samples through highthroughput sequencing and culture dependent. Forty-nine strains were isolated and identified as 9 genera and 15 species,with 11 of which were lactic acid bacteria and 4 as other bacteria. Particularly, Leuconostoc bacteria turned out to be the majority of the isolated microbe. On the other hand, data from high-throughput sequencing revealed that more than 50% of the bacteria belong to lactic acid bacteria(LAB) in H1 and H3 samples. A total of two hundred and twelve species were found by high-throughput sequencing and sixty-six of them belonged to LAB. High-throughput sequencing indicated that stinky tofu flora composition is very complicated and no one species was identified as the dominant strain among all the stinky tofu in this study. Lactobacillus sakei is isolated from each sample and takes a relatively high percentage in this specific niche.
引文
[1]GU J S, LIU T J, SADIQ F A, et al. Biogenic amines content and assessment of bacterial and fungal diversity in stinky tofu-A traditional fermented soy curd[J]. LWT-Food Science and Technology,2018, 88(Supplement C):26-34.
[2]卢义伯,潘超,祝义亮.臭豆腐发酵菌种的筛选与鉴定[J].食品科学, 2007, 28(6):246-249.
[3]CHAO, S H, TOMII Y, WATANABE K, et al.Diversity of lactic acid bacteria in fermented brines used to make stinky tofu[J]. International Journal of Food Microbiology, 2008, 123(1/2):134-141.
[4]LIU T J, LI Y, SADIQ F A, et al. Predominant yeasts in Chinese traditional sourdough and their influence on aroma formation in Chinese steamed bread[J]. Food Chemistry, 2018, 242:404-411.
[5]曲巍,张智,马建章,等.高通量测序研究益生菌对小鼠肠道菌群的影响[J].食品科学, 2017, 38(1):214-219.
[6]叶雷,闫亚丽,陈庆森,等.高通量测序技术在肠道微生物宏基因组学研究中的应用[J].中国食品学报, 2016, 16(7):216-223.
[7]张和平,郑艺.高通量测序技术在乳制品研究中的应用[J].中国食品学报, 2015, 15(3):1-7.
[8]LIU T J, LI Y, CHEN Y Y, et al. Prevalence and diversity of lactic acid bacteria in Chinese traditional sourdough revealed by culture dependent and pyrosequencing approaches[J]. LWT-Food Science and Technology, 2016, 68:91-97.
[9]陈泽斌,李冰,徐胜光,等.高通量测序分析食用玫瑰花瓣内生细菌多样性[J].中国食品学报, 2017,17(9):227-235.
[10]LEE S F, HUANG W H, LIU Y C, et al. Alkaline fermented foods-stinky tofu:screening and identification of fermenting microflora for stinky brine[J].Food Science, Taiwan, 1999, 26(4):384-394.
[11]CHAO, S H, TOMII Y, SASAMOTO M, et al.Lactobacillus capillatus sp nov., a motile bacterium isolated from stinky tofu brine[J]. International Journal of Systematic and Evolutionary Microbiology,2008, 58(11):2555-2559.
[12]CUEVA C, MORENO-ARRIBAS M V, BARTOLOME B, et al, Antibiosis of vineyard ecosystem fungi against food-borne microorganisms[J]. Research In Microbiology, 2011, 162(10):1043-1051.
[13]FRANZETTI L, SCARPELLINI M. Characterisation of Pseudomonas spp. isolated from foods[J]. Annals of Microbiology, 2007, 57(1):39-47.
[14]赵国忠,谢小芳,邸海波,等.臭豆腐卤水微生物群落及其接种发酵[J].中国酿造, 2017, 36(2):21-24.
[15]CAI X H, MA J, WEI D Z, et al. Functional expression of a novel alkaline-adapted lipase of Bacillus amyloliquefaciens from stinky tofu brine and development of immobilized enzyme for biodiesel production[J]. Antonie Van Leeuwenhoek International Journal of General And Molecular Microbiology,2014, 106(5):1049-1060.
[16]何理,吴晖.市售臭豆腐中菌种的分离及生化鉴定[J].现代食品科技, 2012, 28(8):952-955, 958.
[17]HE S H, MA Y, ZHOU S J, et al. Antioxidant activities of fermented soybean prepared with Bacillus subtilis[J]. Asian Journal of Chemistry, 2013,25(18):10565-10568.
[18]伍蕊,沈宁,朱红,等.克氏库克菌血流感染1例并文献复习[J].中国感染与化疗杂志, 2015, 15(4):380-382.
[19]贺静,谢靓,曾玉伦,等. 454高通量技术比较两种不同臭豆腐卤水中微生物多样性[J].微生物学杂志,2017, 37(4):46-52.
[20]LIU S N, HAN Y, ZHOU Z J. Lactic acid bacteria in traditional fermented Chinese foods[J]. Food Research International, 2011, 44(3):643-651.
[21]SUN G P, ZHANG X J, WANG Y, et al. The investigation of bacteria diversity in stinky tofu brine[J]. Modern Food Science and Technology, 2010,26(10):1087-1091.
[22]LIU C J, GONG F M, LI X R, et al. Natural populations of lactic acid bacteria in douchi from Yunnan Province, China[J]. Journal of Zhejiang University-Science B, 2012, 13(4):298-306.
[2]卢义伯,潘超,祝义亮.臭豆腐发酵菌种的筛选与鉴定[J].食品科学, 2007, 28(6):246-249.
[3]CHAO, S H, TOMII Y, WATANABE K, et al.Diversity of lactic acid bacteria in fermented brines used to make stinky tofu[J]. International Journal of Food Microbiology, 2008, 123(1/2):134-141.
[4]LIU T J, LI Y, SADIQ F A, et al. Predominant yeasts in Chinese traditional sourdough and their influence on aroma formation in Chinese steamed bread[J]. Food Chemistry, 2018, 242:404-411.
[5]曲巍,张智,马建章,等.高通量测序研究益生菌对小鼠肠道菌群的影响[J].食品科学, 2017, 38(1):214-219.
[6]叶雷,闫亚丽,陈庆森,等.高通量测序技术在肠道微生物宏基因组学研究中的应用[J].中国食品学报, 2016, 16(7):216-223.
[7]张和平,郑艺.高通量测序技术在乳制品研究中的应用[J].中国食品学报, 2015, 15(3):1-7.
[8]LIU T J, LI Y, CHEN Y Y, et al. Prevalence and diversity of lactic acid bacteria in Chinese traditional sourdough revealed by culture dependent and pyrosequencing approaches[J]. LWT-Food Science and Technology, 2016, 68:91-97.
[9]陈泽斌,李冰,徐胜光,等.高通量测序分析食用玫瑰花瓣内生细菌多样性[J].中国食品学报, 2017,17(9):227-235.
[10]LEE S F, HUANG W H, LIU Y C, et al. Alkaline fermented foods-stinky tofu:screening and identification of fermenting microflora for stinky brine[J].Food Science, Taiwan, 1999, 26(4):384-394.
[11]CHAO, S H, TOMII Y, SASAMOTO M, et al.Lactobacillus capillatus sp nov., a motile bacterium isolated from stinky tofu brine[J]. International Journal of Systematic and Evolutionary Microbiology,2008, 58(11):2555-2559.
[12]CUEVA C, MORENO-ARRIBAS M V, BARTOLOME B, et al, Antibiosis of vineyard ecosystem fungi against food-borne microorganisms[J]. Research In Microbiology, 2011, 162(10):1043-1051.
[13]FRANZETTI L, SCARPELLINI M. Characterisation of Pseudomonas spp. isolated from foods[J]. Annals of Microbiology, 2007, 57(1):39-47.
[14]赵国忠,谢小芳,邸海波,等.臭豆腐卤水微生物群落及其接种发酵[J].中国酿造, 2017, 36(2):21-24.
[15]CAI X H, MA J, WEI D Z, et al. Functional expression of a novel alkaline-adapted lipase of Bacillus amyloliquefaciens from stinky tofu brine and development of immobilized enzyme for biodiesel production[J]. Antonie Van Leeuwenhoek International Journal of General And Molecular Microbiology,2014, 106(5):1049-1060.
[16]何理,吴晖.市售臭豆腐中菌种的分离及生化鉴定[J].现代食品科技, 2012, 28(8):952-955, 958.
[17]HE S H, MA Y, ZHOU S J, et al. Antioxidant activities of fermented soybean prepared with Bacillus subtilis[J]. Asian Journal of Chemistry, 2013,25(18):10565-10568.
[18]伍蕊,沈宁,朱红,等.克氏库克菌血流感染1例并文献复习[J].中国感染与化疗杂志, 2015, 15(4):380-382.
[19]贺静,谢靓,曾玉伦,等. 454高通量技术比较两种不同臭豆腐卤水中微生物多样性[J].微生物学杂志,2017, 37(4):46-52.
[20]LIU S N, HAN Y, ZHOU Z J. Lactic acid bacteria in traditional fermented Chinese foods[J]. Food Research International, 2011, 44(3):643-651.
[21]SUN G P, ZHANG X J, WANG Y, et al. The investigation of bacteria diversity in stinky tofu brine[J]. Modern Food Science and Technology, 2010,26(10):1087-1091.
[22]LIU C J, GONG F M, LI X R, et al. Natural populations of lactic acid bacteria in douchi from Yunnan Province, China[J]. Journal of Zhejiang University-Science B, 2012, 13(4):298-306.